Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

HRM Accuracy and Limitations as a Species Typing Tool for Leishmania Parasites

Version 1 : Received: 22 August 2023 / Approved: 23 August 2023 / Online: 24 August 2023 (09:48:48 CEST)

A peer-reviewed article of this Preprint also exists.

Filgueira, C.P.B.; Pitta-Pereira, D.; Cantanhêde, L.M.; Ferreira, G.E.M.; Dos Reis, S.; Cupolillo, E.; Moreira, O.C.; Britto, C.; Boité, M.C. HRM Accuracy and Limitations as a Species Typing Tool for Leishmania Parasites. Int. J. Mol. Sci. 2023, 24, 14784. Filgueira, C.P.B.; Pitta-Pereira, D.; Cantanhêde, L.M.; Ferreira, G.E.M.; Dos Reis, S.; Cupolillo, E.; Moreira, O.C.; Britto, C.; Boité, M.C. HRM Accuracy and Limitations as a Species Typing Tool for Leishmania Parasites. Int. J. Mol. Sci. 2023, 24, 14784.

Abstract

High Resolution Melting Analysis (HRM) has been pointed as a suitable alternative method to detect and identify Leishmania species. Herein, we aimed to evaluate the sensitivity, specificity, accuracy, and limitations of a HSP70-HRM protocol both as a diagnostic scheme applied in clinical samples and as a species typing tool for laboratory research and reference services. Our data reveal the pronounced species-typing potential of the HSP70-HRM in DNA from cultured parasites. For clinical samples, however, we advise caution due to the parasite load-dependent accuracy. In the light of these findings and considering the importance of parasite load determination for clinical and research purposes we recommend the integration of the presented typing scheme and the previously published Leishmania quantifying approach as combined tools for clinicians, surveillance, and research.

Keywords

HRM; Leishmania; Americas; leishmaniasis; qPCR; parasite typing; HSP70; diagnosis.

Subject

Biology and Life Sciences, Parasitology

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